牛血清白蛋白与光谱探针相互作用机理的研究

在酸性条件下研究了考马斯亮蓝光谱探针与牛血清白蛋白结合物的光学性质,探讨了考马斯亮蓝染料和蛋白质的相互作用机理。研究结果表明染料考马斯亮蓝的疏水作用和亲水作用的协调作用以及与生物大分子蛋白质具有相反电荷的静电作用,是光谱探针与生物大分子形成大分子聚集体的必要因素。     

Molecular simulation of the interaction between novel type rhodanine derivative probe and bovine serum albumin

The interaction between 3-(4′-methylphenyl)-5-(4′-methyl-2′-sulfophenylazo) rhodanine (M4MRASP) and bovine serum albumin (BSA) was studied by using spectrofluorimetry. It was shown in fluorescence spectrums that the quenching mechanism of BSA by M4MRASP was a static quenching. Meanwhile, the binding constant and binding site numbers were calculated. The action distance (r = 8.03 nm) and energy transfer efficiency (E = 0.12) between donor (BSA) and acceptor (M4MRASP) were obtained according to the theory of Förster non-radiation energy transfer. The effect of M4MRASP on the conformation of BSA was further analyzed by using synchronous fluorescence spectrometry. A new model of the interaction between small organic molecule and biomacromolecule was established. The results offered a reference for the studies on the biological effects and action mechanism of small molecule with protein.     

Thermal stability of bovine serum albumin DSC study

A calorimetric study of thermal denaturation of bovine serum albumin in aqueous solutions has shown essential differences in stability of fatty acid containing and defatted albumin. The first one shows a single endotherm peak in DSC curve near 69°C with enthalpy change about 1000 kJ mol^-1. Defated albumin melts in two different temperature ranges: near 56 and 69°C with enthalpy changes about 300 and 200 kJ mol^-1 respectively. Deconvolution analysis shows that the single endotherm is well approximated as the sum of three independent two-state transitions. Two transitions of bimodal DSC curve for defatted albumin are not of a two-state type. This molecule melts probably as two structurally independent parts. This revised version was published online in July 2006 with corrections to the Cover Date.     

Electrostatic interaction between chitosan-modified latex particles and bovine serum albumin

Electrostatic interaction between poly(methyl methacrylate) latex particles with different levels of chitosan modification and bovine serum albumin (BSA) was investigated. The critical flocculation concentration is in the range 5–15 nmol dm⁻³ for these latex products toward added BSA. A series of isothermal equilibrium adsorption experiments shows that the adsorption process is divided into two distinct intervals. Adsorption of BSA on latex particles in intervals I and II is primarily controlled by charge neutralization and hydrophobic interaction, respectively. Intervals I and II can be reasonably described by an empirical parabola equation and the Langmuir isotherm model, respectively. The maximum amount of BSA adsorbed per unit weight of polymer particles was observed at pH ≅ 5. A maximum elution yield of about 80% can be achieved using NaSCN as the elution electrolyte, and NaSCN is more effective in inducing desorption of BSA from the particle surface than NaCl. The chitosan content has very little effect on the interaction between latex particles and BSA. By contrast, the influence of the content of 2,2′-azobis(2-amidinopropane) dihydrochloride, a cationic initiator used in preparing the chitosan-modified latex products, on the BSA adsorption process is significant. Received: 26 March 1999 Accepted in revised form: 3 June 1999     

Interaction of magnolol with bovine serum albumin: a fluorescence-quenching study

The interaction of magnolol with bovine serum albumin(BSA) was studied using fluorescence spectroscopy under physiological conditions. The binding constants, K, and the ratio of quantum yields of protein fluorescence for complex and free protein, f, at 298 K, 304 K, and 310 K were obtained; the values were 6.799×10⁵ L mol^–1, 5.541×10⁵ L mol^–1, and 4.344×10⁵ L mol^–1 and 0.17, 0.30, and 0.34, respectively. The standard enthalpy change (H°) and the standard entropy change (S°) were calculated to be –28.53 kJ mol^–1 and 15.88 J mol^–1 K^–1, which indicated that hydrophobic forces played major role in the interaction of magnolol and BSA. The binding average distance between magnolol and BSA (4.32 nm) was obtained on the basis of the theory of Förster energy transfer.     

溴甲酚绿分光光度法测定牛血清白蛋白

在pH 3.3的Britton-Robinson (B-R)缓冲溶液中, 对溴甲酚绿(BCG)与牛血清白蛋白(BSA)相互作用的吸收光谱进行了初步研究. 结果表明: BCG与BSA作用在室温下能迅速结合成复合物, 并且随着BSA的浓度增大, 在444 nm处的吸收峰降低, 618 nm处吸收峰升高并红移至628 nm. 在此波长下测定其复合物的吸光度, 其吸光度的增加值(ΔA)与BSA的质量浓度在8～260 μg/mL范围内呈良好的线性关系(r=0.9996), 检出限为4 μg/mL. 该方法应用于鲜奶粉和液态纯牛奶样品中总蛋白的测定, 回收率分别为92.7%, 95.5%, 结果与考马斯亮蓝G250法基本一致.     

Linear sweep voltammetric determination of protein based on its interaction with Alizarin Red S

In this paper, the electrochemical behavior of the interaction of Alizarin Red S (ARS) with bovine serum albumin (BSA) was investigated on the hanging mercury drop electrode (HMDE). In the acidic solution (pH 4.2), ARS can be easily reduced on the HMDE and it has a well-defined polarographic wave at −0.29 V (SCE). On addition of BSA or human serum albumin (HAS) into the ARS solution, the reduction peak current of ARS decreases without the movement of the peak potential and the appearance of new peaks. The study shows that a new electrochemically non-active complex is formed via intercalation of ARS with BSA or HSA, which can not be reduced on the Hg electrode. The decrease of reductive peak current of ARS is proportional to BSA and HSA concentration in the range of 2.0–60 and 2.0–40 mg l⁻¹, respectively. The detection limit of BSA and HSA is 1.0-mg l⁻¹. The analytical results of human serum and urine samples by this method were in good agreement with the Coomassie brilliant blue G-250 assay. The binding number and the binding interaction mechanism are also discussed.     

6-苄氨基嘌呤与牛血清白蛋白作用荧光特性的研究

应用荧光光谱法研究了6-苄氨基嘌呤（6-BA）与牛血清白蛋白（BSA）相互作用的荧光特性。测得了6-BA与BSA在10、27、40℃温度下的结合常数KA为：0.21×10^5、1.37×10^5、5.53×10^5L/mol,结合位点数n为：1.0、1.2、1.3。6-BA对BSA内源荧光的猝灭机理主要为静态猝灭,6-BA主要以疏水作用与BSA相互作用,BSA的荧光主要源于色氨酸残基,6-BA对BSA的构象有影响。     

CE determination of quinolones in the presence of bovine serum albumin

This article describes the influence of bovine serum albumin (BSA) as an additive on the capillary electrophoresis–potential gradient determination of five quinolones, enoxacin, norfloxacin, ofloxacin, fleroxacin, and pazufloxacin. With 10 mg/L of BSA present in the buffer of 30 mM Tris and 3 mM phosphoric acid at pH 9, the detection limits of the five quinolones were in the range of 0.24–0.68 mg/L, i. e. 5.8–16.5‐fold lower than those obtained with the buffer devoid of BSA, and the analysis time was shortened. We suggest that the inner wall‐adsorbed BSA suppresses the adsorption of quinolones and simultaneously enhances the electroosmotic flow rate. Our experiments indicated that adopting the potential gradient detection technique could eliminate the interference of the UV‐active proteins on the detection of quinolones that would occur with conventional optical detection, and therefore offer high detection sensitivity. As a demonstration, the method was applied to the determination of QNs in fortified chicken muscle sample with satisfactory results.     

二氢氯噻与牛血清白蛋白的相互作用特征

利用荧光技术研究了在生理酸度条件下,二氢氯噻与牛血清白蛋白相互作用,发现二氢氯噻对牛血清白蛋白有较强的荧光猝灭作用,用Stern-Volmer和Line weaver-Burk方程处理荧光猝灭数据,得到了反应的结合常数、结合热力学性质等参数。根据热力学参数确定了该药物与血清白蛋白之间的作用力类型,在此基础上依据福斯特F rster非辐射能量转移理论探讨了二氢氯噻与BSA相互结合时其供体-受体间的距离。与人血清蛋白[1]比较,牛血清白蛋白与二氢氯噻的结合较弱,体现了二氢氯噻与血清蛋白结合的动物间的差异性。同时考察了中药活性成分和金属离子对结合的影响,结果显示甘草次酸对结合的影响较大,提示同时给药时应该注意它们之间的血清药物相互作用。     

金丝桃苷与牛血清白蛋白相互作用的研究

采用荧光共振能量转移法研究了金丝桃苷与牛血清白蛋白的相互作用.金丝桃苷对牛血清白蛋白(BSA)的荧光猝灭类型是静态猝灭,25℃时的结合位点数为0.5451.并依据F(o)ster非辐射能量转移理论,研究了给体(牛血清白蛋白)--受体(金丝桃苷)间的结合距离R.和能量转移效率E分别为2.04nm和0.66.同时,采用同步...     

Electrochemistry of the interaction of furazolidone and bovine serum albumin

The electrochemical behavior of the interaction of furazolidone (Fu) with bovine serum albumin (BSA) was investigated by cyclic voltammetry and differential pulse voltammetry at a glassy carbon electrode. Fu shows an irreversible reduction at − 0.34 V in pH 4.0 Britton–Robinson buffer (B–R) buffer–10% DMF solution. After the addition of BSA into the Fu solution, the reductive peak currents decreased without any significant shift of the peak potential and the appearance of new peaks. The electrochemical parameters of the interaction system were calculated in the absence and presence of BSA. This electrochemical method was further applied to the determination of BSA samples and the results were in good agreement with the traditional cellulose acetate electrophoresis. The linear dynamic range was between 10.0 and 80.0 mg l^− 1. The detection limit was 7.6 mg l^− 1 and the recoveries were obtained from 97.0% to 104.0%.     

Rhenium(I)-based fluorescence resonance energy transfer probe for conformational changes of bovine serum albumin

Protein binding properties of fac-rhenium(I) complexes with general structure [Re(CO)₃(N-N)L]PF₆, where N-N = 4,4′-dinanoyl-2,2-bipyridine and L = py-3-COOH (1a) and py-3-CONH₂ (1b) with bovine serum albumin (BSA) were investigated at physiological pH (7.4) using UV-visible absorption and fluorescence spectral study, excited state lifetime measurement and circular dichroism (CD). The results observed from fluorescence spectra reveal the energy transfer from BSA to Re(I) complex, and the distance r between donor (BSA) and acceptor (Re(I) complex) is 3.05 nm and 2.16 nm for 1a and 1b respectively according to Forster's non-radiative energy transfer theory. CD results show that the binding of Re(I) complex could induce the conformational change with the loss of α-helicity.     

秋水仙碱与牛血清白蛋白相互作用的电化学研究

以电化学方法对秋水仙碱与牛血清白蛋白的相互作用进行了研究。在0.3 mol/L H2SO4底液中,秋水仙碱在玻碳电极上产生一不可逆的氧化峰,峰电位为1.18 V(vs.SCE),加入表面活性剂四丁基氯化铵后,秋水仙碱的峰信号得到明显提高。在上述条件下,加入BSA后秋水仙碱的氧化峰电位正移,峰电流下降,峰电流下降值与BSA加入的浓度在1.5×10-7～2×10-6mol/L(r=0.9978)范围内有良好的线性关系,检出限达4.0×10-8mol/L。进一步探讨了秋水仙碱与BSA的结合数和结合常数,得到结合数为1,结合常数为2.40×105L/mol。     

基于分子印迹技术的牛血清白蛋白电位式传感器的研究

以牛血清白蛋白(BSA)为模板,合成了分子印迹聚合物凝胶(MIP),进而制得了以聚氯乙烯(PVC)为支撑膜的BSA电位式传感器。采用直接电位法测得BSA在浓度0.1～1.0 mg/mL范围内与电位呈线性关系。此电极制作简单,可用于BSA的测定。     

Study on the interaction between clarithromycin and bovine serum albumin in the imitated physiology solution

The interaction between clarithromycin (CAM) and bovine serum albumin (BSA) was investigated using linear-sweep voltammetry in pH 7.4 phosphate buffer solution where CAM caused two irreversible reduction waves P2 and P3 on mercury electrode. The study showed that the formation constant and formation ratio for the interaction between CAM and BSA were 1.51×1012 and 3:1 for P2,4.53×105 and 1:1 for P3, respectively. The ion strength enhanced the hydrophobic interaction between CAM and BSA.     

A kind of modified bovine serum albumin with great potential for applying in gene delivery

Bovine serum albumin(BSA) was modified through a facile synthesis method to increase its isoelectric point(pI) from 4.8 to 6.0.When pH is higher than 6.0,the protein shows a negative surface charge,on the contrary,the protein is positively charged.In this study,the charge-reversal modified BSA(crBSA) was utilized to assemble with the binary complexes of pDNA/poly(vinylpyrrolidone)-graft-poly(2-dimethylaminoethyl methacrylate)(pDNA/PVP-g-PDMAEMA) to shield the excess positive charges of complexes at physiological pH(pH 7.4).When the complex coated with crBSA located in the environment at endosomal pH(pH 5.0),the charge-reversal of crBSA led to the deviation of crBSA from polyplex by electrostatic repulsion,which would benefit the transfection of the target gene.The crBSA shows great potential for improving the transfection efficiency of pDNA/PVP-g-PDMAEMA.     

2,4-二硝基苯胺与牛血清白蛋白相互作用的研究

通过荧光和紫外光谱法研究了2,4-二硝基苯胺同牛血清白蛋白(BSA)的相互作用. 2,4-二硝基苯胺对牛血清白蛋白的内源荧光具有强烈的猝灭作用. 二者之间形成不发荧光的复合物是导致荧光猝灭的主要原因. 计算了其结合常数和结合位点数. 紫外光谱法进一步证明了其猝灭机理为静态猝灭. 根据能量转移理论计算了作用距离(3.13 nm). 同步荧光的结果表明2,4-二硝基苯胺的存在改变了牛血清白蛋白的分子构象.     

卡络磺钠与牛血清蛋白荧光光谱的研究

采用荧光光谱法研究了卡络磺钠CSS (Carbazochrome Sodium Sulfonate)与牛血清蛋白(BSA)结合反应的特征, 测定了结合常数(K=1.32×105 L/mol) 和结合位点数(n=1.28). 依据Foster非辐射能量转移理论, 确定了给体-受体间的结合距离(r=4.896 nm)和能量转移效率, 采用同步荧光技术考察了CSS对BSA构象的影响.